{"title":"喜马拉雅山濒危药用芳香植物当归的体外繁殖和次生代谢物生产","authors":"Deepika Negi, Manisha Thakur, Bhupender Dutt, Rohit Sharma","doi":"10.1007/s11240-024-02825-2","DOIUrl":null,"url":null,"abstract":"<p>This work presents an efficient one-step procedure for in vitro propagation in <i>Angelica glauca</i> using rhizome buds and production of secondary metabolites. A maximum of 94% of buds were established in vitro on medium supplemented with 0.3 mg/L Benzyl adenine (BA) and 0.1 mg/L Gibberellic acid (GA<sub>3</sub>). After the fifth sub-culture, the proliferating shoots from the rhizome buds displayed the maximum proliferation (1:15), rooted on Murashige and Skoog (MS) medium supplemented with 1.0 mg/L BA and 0.2 mg/L α-naphthalene acetic acid (NAA). After being transferred to pots with soil:cocopeat (1:1) for hardening, shoots with enlarged rhizomes demonstrated 60% survival after a month in the polyhouse. For secondary metabolite production, callus was induced from in vivo roots on MS medium supplemented with 0.5 mg/L Kinetin (Kin) and 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) under dark incubation and after 1 year its suspension culture showed the existence of 206 compounds. The gas chromatography-mass spectrometry (GC–MS) analysis results revealed that extracts predominantly contain compounds from different classes such as esters, ethers, and <i>N</i>-heterocyclic pyrrolo pyridazine, fatty acids and mono and sesquiterpenes with varying concentrations. On elicitation with 0.5, 1.0 and 1.5 mM Methyl jasmonate (MeJA) the callus cultures depicted varying concentration of monoterpene such as <span>d</span>-limonene, <i>trans</i> and <i>cis</i>-ligustilide, a marker compound of <i>A. glauca</i> essential oil, fatty acids and ethers. Sucrose treatment at 1, 3 and 5% revealed the presence of various unsaturated fatty acids, hydrocarbon, ethers, sesquiterpenes β-farnesene, α-copaene, and carotenoid rhodopin. Addition of growth regulators (2,4-D and Kin) revealed the presence of furfural and its derivatives, benzoic acids and esters.</p>","PeriodicalId":20219,"journal":{"name":"Plant Cell, Tissue and Organ Culture","volume":"128 1","pages":""},"PeriodicalIF":2.3000,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"In vitro propagation and secondary metabolites production of Angelica glauca Edgew: a threatened medicinal and aromatic herb of the Himalayas\",\"authors\":\"Deepika Negi, Manisha Thakur, Bhupender Dutt, Rohit Sharma\",\"doi\":\"10.1007/s11240-024-02825-2\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>This work presents an efficient one-step procedure for in vitro propagation in <i>Angelica glauca</i> using rhizome buds and production of secondary metabolites. A maximum of 94% of buds were established in vitro on medium supplemented with 0.3 mg/L Benzyl adenine (BA) and 0.1 mg/L Gibberellic acid (GA<sub>3</sub>). After the fifth sub-culture, the proliferating shoots from the rhizome buds displayed the maximum proliferation (1:15), rooted on Murashige and Skoog (MS) medium supplemented with 1.0 mg/L BA and 0.2 mg/L α-naphthalene acetic acid (NAA). After being transferred to pots with soil:cocopeat (1:1) for hardening, shoots with enlarged rhizomes demonstrated 60% survival after a month in the polyhouse. For secondary metabolite production, callus was induced from in vivo roots on MS medium supplemented with 0.5 mg/L Kinetin (Kin) and 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) under dark incubation and after 1 year its suspension culture showed the existence of 206 compounds. The gas chromatography-mass spectrometry (GC–MS) analysis results revealed that extracts predominantly contain compounds from different classes such as esters, ethers, and <i>N</i>-heterocyclic pyrrolo pyridazine, fatty acids and mono and sesquiterpenes with varying concentrations. On elicitation with 0.5, 1.0 and 1.5 mM Methyl jasmonate (MeJA) the callus cultures depicted varying concentration of monoterpene such as <span>d</span>-limonene, <i>trans</i> and <i>cis</i>-ligustilide, a marker compound of <i>A. glauca</i> essential oil, fatty acids and ethers. Sucrose treatment at 1, 3 and 5% revealed the presence of various unsaturated fatty acids, hydrocarbon, ethers, sesquiterpenes β-farnesene, α-copaene, and carotenoid rhodopin. 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引用次数: 0
摘要
本研究提出了一种利用根茎芽体外繁殖当归并生产次生代谢物的高效一步法。在添加了 0.3 毫克/升苄基腺嘌呤(BA)和 0.1 毫克/升赤霉素(GA3)的培养基上,最多有 94% 的芽在离体培养中成活。第五次亚培养后,根茎芽中的增殖芽在添加了 1.0 mg/L BA 和 0.2 mg/L α-萘乙酸(NAA)的 Murashige and Skoog(MS)培养基上生根,增殖率最高(1:15)。将其转移到装有土壤与椰糠(1:1)的花盆中进行硬化处理后,在温室中生长一个月后,根茎增大的嫩芽存活率达到 60%。为了生产次生代谢物,在添加了 0.5 毫克/升酮素(Kin)和 2.0 毫克/升 2,4-二氯苯氧乙酸(2,4-D)的 MS 培养基上,在黑暗培养条件下从活体根部诱导出胼胝体。气相色谱-质谱(GC-MS)分析结果表明,萃取物主要含有不同类别的化合物,如酯、醚、N-杂环吡咯哒嗪、脂肪酸、单萜和倍半萜,浓度各不相同。用 0.5、1.0 和 1.5 mM 的茉莉酸甲酯(MeJA)诱导时,胼胝体培养物显示出不同浓度的单萜烯,如 d-柠檬烯、反式和顺式藁本内酯(A. glauca 精油的标记化合物)、脂肪酸和醚。蔗糖浓度为 1%、3% 和 5%时,各种不饱和脂肪酸、碳氢化合物、醚类、倍半萜烯 β-法呢烯、α-罂粟碱和类胡萝卜素荷包牡丹碱都会出现。加入生长调节剂(2,4-D 和 Kin)后,发现存在糠醛及其衍生物、苯甲酸和酯类。
In vitro propagation and secondary metabolites production of Angelica glauca Edgew: a threatened medicinal and aromatic herb of the Himalayas
This work presents an efficient one-step procedure for in vitro propagation in Angelica glauca using rhizome buds and production of secondary metabolites. A maximum of 94% of buds were established in vitro on medium supplemented with 0.3 mg/L Benzyl adenine (BA) and 0.1 mg/L Gibberellic acid (GA3). After the fifth sub-culture, the proliferating shoots from the rhizome buds displayed the maximum proliferation (1:15), rooted on Murashige and Skoog (MS) medium supplemented with 1.0 mg/L BA and 0.2 mg/L α-naphthalene acetic acid (NAA). After being transferred to pots with soil:cocopeat (1:1) for hardening, shoots with enlarged rhizomes demonstrated 60% survival after a month in the polyhouse. For secondary metabolite production, callus was induced from in vivo roots on MS medium supplemented with 0.5 mg/L Kinetin (Kin) and 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) under dark incubation and after 1 year its suspension culture showed the existence of 206 compounds. The gas chromatography-mass spectrometry (GC–MS) analysis results revealed that extracts predominantly contain compounds from different classes such as esters, ethers, and N-heterocyclic pyrrolo pyridazine, fatty acids and mono and sesquiterpenes with varying concentrations. On elicitation with 0.5, 1.0 and 1.5 mM Methyl jasmonate (MeJA) the callus cultures depicted varying concentration of monoterpene such as d-limonene, trans and cis-ligustilide, a marker compound of A. glauca essential oil, fatty acids and ethers. Sucrose treatment at 1, 3 and 5% revealed the presence of various unsaturated fatty acids, hydrocarbon, ethers, sesquiterpenes β-farnesene, α-copaene, and carotenoid rhodopin. Addition of growth regulators (2,4-D and Kin) revealed the presence of furfural and its derivatives, benzoic acids and esters.
期刊介绍:
This journal highlights the myriad breakthrough technologies and discoveries in plant biology and biotechnology. Plant Cell, Tissue and Organ Culture (PCTOC: Journal of Plant Biotechnology) details high-throughput analysis of gene function and expression, gene silencing and overexpression analyses, RNAi, siRNA, and miRNA studies, and much more. It examines the transcriptional and/or translational events involved in gene regulation as well as those molecular controls involved in morphogenesis of plant cells and tissues.
The journal also covers practical and applied plant biotechnology, including regeneration, organogenesis and somatic embryogenesis, gene transfer, gene flow, secondary metabolites, metabolic engineering, and impact of transgene(s) dissemination into managed and unmanaged plant systems.